Prepare Peptone Yeast Extract (PYE) nutrient medium selectively culture Caulobactercrescentus. This protocol describes the preparation of 500 - 1000 mL of medium, adapted from both DSMZ 1462: CAULOBACTER PYE MEDIUM and a protocol provided by our advisor, Beth.
Procedure
Follow the same procedure as LB Medium Recipe, using the following reagents instead:
Version 1 (from standard)
Component
Amount (for 500 mL)
Final concentration
Peptone
1 g
2 g/L
Yeast extract
0.5 g
1 g/L
CaCa₂·2H₂O (Calcium chloride dihydrate)
0.05 g
0.1 g/L
MgSO₄·2H₂O (Magnesium sulfate heptahydrate)
0.1 g
0.2 g/L
Optional: Agar (for solid medium)
6 g
1.2%
Version 2 (from manuscript)
0.3% peptone, 0.2% yeast extract, 1.5 mM MgSO4, 1 mM CaCl2, 1.2% agar for solid medium
Component
Amount (for 1000 mL)
Final concentration
Peptone
3.00 g
3 g/L
Yeast extract
2.00 g
2 g/L
CaCa₂·2H₂O (Calcium chloride dihydrate)
219.08 mg
0.219 g/L or 1 mM
MgSO₄·2H₂O (Magnesium sulfate heptahydrate)
369.71 mg
0.370 g/L or 1.5 mM
Optional: Agar (for solid medium)
12.00 g
1.2%
Note that the concentration of agar can be increase to 1.5% for a denser gel.